Diet supplementation with fruits and vegetables has an impact of preventing and reversing defects associated with an aging subject. Nutritional regimes that involve dietary supplementations with berryfruit (e.g., Vaccinium berries such as cranberries and blueberries) reverse and/or forestall both motor and cognitive changes associated with aging. For example, blueberry supplementation prevents cognitive behavioral deficits in mice having increased amyloid β-peptide production with APP/PS-1 mutations. (Joseph J A, et al., 2003. Nutr. Neurosci., 6; 153-162). The beneficial effects of berryfruits involve direct and indirect actions against oxidative stressors. As such, there is a need to identify compounds in berryfruit that are effective in altering oxidative mediated changes in motor and cognitive function.
Trans-3,5,4′-trihydroxystilbene (hereinafter referred to as resveratrol) has been identified as having a plurality of anti-aging properties due to its robust anti-oxidant activity. In vitro experiments have shown that resveratrol is an effective free radical scavenger and inhibits low density lipoprotein oxidation (Brito P, et al., 2002. Free Radic Res. 36(6):621-633.). Other stilbenoids such as pinostilbene, desoxyrhapontigenin, pterostilbene, resveratrol trimethylether, and piceatannol, have varying degree of biological activity and effectiveness for lowering lipid levels by activating a nuclear receptor, peroxisome proliferators activated receptor alpha isoform.
Trans-3,5-dimethoxy-4′-hydroxystilbene (hereinafter referred to as pterostilbene), a natural methylether analog of resveratrol, had been demonstrated to have antioxidant activity similar to that of resveratrol (Rimando et al., 2002. J. Agric. Food Chem. 50:3453-3457; Stivala et al., 2001. J. Biol. Chem. 276(25):22586-22594.) Pterostilbene is present in some small fruits such as grapes (Adrian et al., 2000. J. Agric. Food Chem. 48:6103-6105) and berries of Vaccinium (Vaccinium ashei Reade and Vaccinium stamineum L.) (Rimando et al., 2004. J. Agric. Food Chem. 52:4713-4719) as well as in woody plants (Maurya et al., 1977, J. Nat. Prod. 47:179-181; Amone et al., 1977. J. Chem. Soc. Perkins Trans. 19:2116-2118). Additionally, a plurality of botanicals contain pterostilbene, including Anogeissus acuminata, Dracaena cochinchinensis, Dracaena loureiri, Guibourtia tessmannii, Pterocarpus macrocarpus, Pterocarpus marsupium, Pterocarpus santalinus, Vaccinium ashei, Vaccinium corymbosum, Vaccinium deliciosum, Vaccinium membranaceum, Vaccinium ovatum, Vaccinium ovalifollum, Vaccinium parviflorum, Vaccinium stamineum, Vaccinium uliginosum, and Vitis vinifrea, Pterostilbene is also found in non-botanical sources such as propolis.
The level of pterostilbene can vary from species of Vaccinium berries. As reported by Rimando et al., 2004. J. Agric. Food Chem. 52:4713-4719, pterostilbene concentrations via cultivars of Vaccinium berries were reported having 99 ng/gm to 520 ng/gm of dry sample. Additionally, as reported by Rimando et al., Acta Hort. (ISHS), 680:137-143, lyophilized berries from Vaccinium nine cultivars species exhibited 0.12 μg to 2.74 μg of pterostilbene per gram of berries. Similarly, blueberry species vary in the amount of pterostilbene concentration. It has been reported that a range of 99 ng to 475 ng of pterostilbene can be derived from one grain of lyophilized blueberries.
Studies have linked the effects of antioxidants with deleterious effects of brain aging and behavior. The combination of antioxidant/anti-inflammatory polyphenolics found in fruits and vegetables in the form of “secondary chemicals” not generally involved in the plant primary metabolism, has exhibited efficacy in preventing these deleterious effects. As such, there is a need to further identify fruit and plants, specifically the compounds that can protect against aging and cognitive defects.
The compound pterostilbene has shown moderate inhibition of cyclooxygenase-1, and weak inhibition of cyclooxygenase-2 to suggest anti-inflammatory activity. (Rimando, et al., 2002. J. Agric. Food Chem. 50: 3453-3457.) In addition, pterostilbene has been identified to activate peroxisome proliferator-activated alpha isoform, (PPARα) a receptor proposed to mediate lowering lipid and glucose levels. Details of the PPARα agonist effect of pterostilbene is disclosed in Rimando et al., 2005. J. Agric. Food Chem. 53:3403-3407 and US2006/005723A1, both incorporated herein by reference.
Pterostilbene has been identified to inhibit colon carcinogenesis. Particularly, pterostilbene has suppressed azoxymethane-induced colonic aberrant crypt foci growth in weanling male F344 rats. Additionally, pterostilbene inhibits the expression of inducible nitric oxide synthase (iNOS) in HT-29 human adenocarcinoma cell lines. (Suh et al., 2007. Clin Cancer Res. 13(1) 350-155.)
In an effort to ameliorate cognitive changes in senescent subjects, there is a need in the art to determine whether a supplementation of pterostilbene to senescent subjects would reverse cognitive and motor deficiencies as compared to unsupplemented subjects.
Muscarinic cholinergic receptors mediate the actions of the neurotransmitter acetylcholine in the central and peripheral nervous systems, gastrointestinal system, heart, endocrine glands, lungs, and other tissues. Five distinct muscarinic receptor subtypes have been identified as m1-m5. The m1 subtype is the predominant subtype found in the cerebral cortex and is believed to be involved in the control of cognitive functions.
Conditions associated with cognitive impairment, such as Alzheimer's disease, are accompanied by loss of acetylcholine in the brain. This is believed to be the result of degeneration of cholinergic neurons in the basal forebrain, which innervate areas of the association cortex, and hippocampus, which is involved in higher cognitive processes.
As such, there is a need to identify compounds that increase acetylcholine signaling effect in the brain. Specifically there is a need to identify compounds as muscarinic agonists that are active at various receptor subtypes in the central and peripheral nervous system.